Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
ISSN: 2319-7706 Volume 3 Number 8 (2014) pp. 185-196
http://www.ijcmas.com
Original Research Article
Role of the Functional Food (Pomegranate-Yoghourt) as Hepatoprotective
Effect on Liver Injured Rats
Hala, M.F. El Din1, Sherif S. Mohamed2* and T.M. El-Messery1
1
Dairy Science &Technology Department, National Research Centre, Egypt
2
Nutrition & Food science Department, National Research Centre, Egypt
*Corresponding author
ABSTRACT
Keywords
Pomegranate
juice,
Yoghourt,
antioxidant
activity,
phenolic
compounds,
Pomegranate juice (PJ) is commonly used in traditional medicine due to its
therapeutic properties according to its high content of total phenolic content (TPC)
and antioxidant activity (AA) which related to the management of many disorders.
So, supplementing fresh buffalo skim milk with (PJ) with different ratios (2, 4, 6, 8,
and 10%) and the textural properties, TPC and AA in the resultant fresh yoghurt
during the cold storage for 12 days were evaluated. As the percentage of PJ was
increased, the TPC and AA were increased, while the addition of PJ up to 6% had
an adverse effect on the textural properties. The effect of pomegranate-yoghurt
against hepatic injury (CC14 treatment) in rats was investigated through
determination of some biochemical parameters in serum and liver in addition to the
histophathological examination of liver under different treatments. Forty adult
albino rats were equally divided into five groups; a negative control (C- ), CC14treated rats (C+ ) and groups that received a dietary yoghurt-PJ in 0, 4, 8% ratios.
Results revealed that pomegranate-yoghurt improved the level of biomarkers scores
of CC14-treated rats, and reduced the histopathological changes in liver with the
elevating of PJ ratio. Consequently, administration of yoghurt supplemented with
PJ is acceptable as functional food alleviates the harmful effect of CC14-induced
liver injury and offers a pleasant and effective route in increasing the TPC and
antioxidant intake in our daily diet.
Introduction
enhances the immune response which will
be in turn increase resistance to immunerelated diseases (Meydani et al 2000).
Yoghurt can be developed with fruit juice
which can be part of a healthy diet (Aswal et
al 2012), which increases the aesthetic value
of the new product as a functional pigment
(Ghadge et al 2008; Coïsson et al 2005).
The challenge is, however, choosing the
right functional nutrients and formulating a
message that works. Yoghurt is a fermented
dairy product, having several health
benefits, so is healthier for consumption
(lourens-Hattingh and Viljoen 2001).
Yoghurt is beneficial to our digestive
system, especially stomach and colon; it
185
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
for 10 min. and cooled to 40O C.
Pomegranate juice (PJ) (pH 4.5) was added
at the rate of 2, 4, 6, 8, 10% immediately
after incubation with 3% starter culture
(local market) to avoid the quick reducing of
pH and transferred into 100 ml plastic
containers, lightly sealed and incubated at
45O C until the complete curd formation,
then stored at refrigerator (6±2O C) for 3, 6,
9, 12 days.
Pomegranate (Punica granatum Linn) is a
potential medicinal plant of family
punicaceae (Heber 2011). Pomegranate juice
was used as a healthful beverage (Basu and
Penugonda 2009), since it is a natural rich
source of polyphenols, flavonoid and other
antioxidant. It could be considered as
functional ingredients for its anti radical
activities. It is good supplement for food and
dietetics (Tomás-Barberán et al 2006; Basu
and Penugonda 2009). Pharmacological
properties of the juice were antiinflammatory (Adams et al 2006),
anticancer (Adams et al 2010) and
protection of hepatoxicity (Kaneria et al
2012) by carbon tetrachloride (Pirinccioglu
et al 2012).
Textural evaluation
Texture profile analysis (TPA) performed on
the yoghurt samples using the double
corporation, Slinfold, W. Sussex, UK. From
the force time curve the following
parameters were evaluated by TPA
according to the definitions by International
Dairy Federation (1991).
Therefore the aim of the study was to
evaluate the textural parameters; total
phenolic
content
and
DPP-radical
scavenging activity of yoghurt supplemented
with pomegranate juice, also, the protective
roles of pomegranate juice against the
hepatoxicity induced by carbontetrachlorid
for rats feeding on yoghurt supplemented
with pomegranate juice.
Hardness: force necessary to attain a given
deformation
Cohesiveness: the extant to which a material
can be deformed before it
rupture.
Springiness: in the rate which the sample
returns to its original shape
when the
deforming
force is removed.
Gumminess: Force needed to disintegrate
the sample to a state ready for
swallowing
Chewiness: work needed to masticate the
sample to state ready for
swallowing
Materials and Methods
Preparation of pomegranate juice
Fresh mature pomegranate fruits were
purchased from local market at Giza. They
were cleaned and cut; arils were manually
separated from the peals and piths. The fruit
juice was extracted using an electric juicer
and then filtered through cotton mesh. Fresh
juice (12.2% total solids) was stored in
freezer.
Total phenolic content (TPC)
The total phenolic content determined by
Folin-Ciocalteu Reagent according to Zheng
et al (2001), and calculated as Gallic acid.
Preparation of yoghurt
Radical scavenging activity (RSA)
Fresh skim buffalo milk was obtained from
dairy department of Agriculture College,
Cairo University. Milk was heated to 90O C
The stable free radical DPPH is a rapid test
186
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
can provide information on the ability of a
food compound contains antioxidants to act
as free radical scavenger's ability or
hydrogen donor's atom, according to
(Xuewu et al., 2007).
Forty adult (male and female) albino
Speaque-Dawley rats with body weights 8090g were obtained from the Animal Home
in National Research Center Egypt. Animals
were kept individually in stainless steal
cages, water was allowed adlib tam, and the
room temp. was adjusted at 25O C.
At the end the experimental period, rats
were fasted overnight and in the morning
blood samples were taken from each rat by
open heart puncture under slight ether
anesthesia, and serum was separated. The
abdomen was opened and the liver was
excised then washed with saline plotted
between sheets of filter paper and weighed.
Aspartate aminotransferase (AST), alanine
aminotransferase (ALT), total and direct
bilirubin, and albumin were estimated
according to Reirtman and Frankel (1957);
Dreper and Hadley (1990); Bartholomev and
Delang (1966) respectively. Reduced
glutathione
(GSH)
and
glutathione
peroxidase (GPX) according Ellman (1959),
and Weiss et al (1980) respectively.
Experimental Design
Histopathology study
Rates divided into 5 groups, 8 for each
group as follow:
After blood collection, all rats were rapidly
sacrificed and the liver of each animal was
dissected, weighted and portion of it was
preserved in 10% buffered neutral formalin
and paraffin embedded. Four sections
(5mictons in thickness) were taken from
each liver tissue, each section being at a
distance at least 500 µ from the proceeding
one sections were stained with haematoxylin
and eosin for the histological examination
according Ross et al (1989).
Design of animal experiment
Animals
Group (1): C- fed on standard died only
(according to AIN 1993)
Group (2): C+ treated with CC14 , fed on the
standard diet only.
Group (3): y0 treated with CC14 , fed on
yoghurt only.
Group (3): y4 treated with CC14 , fed on
yoghurt with 4% PJ.
Statistical analysis
Data of the textural analysis were
statistically analyzed using GLM procedures
of the SAS (SAS 1996). Means were
separated by using Duncan s multiple rage
test (Duncan 1955).
Group (3): y8 treated with CC14 , fed on
yoghurt with 8% PJ.
Rats
were
administered
(carbon
tetrachloride, Sigma) by back subcutaneous
injection (0.5 ml of 1:1 mixture of CC14 and
olive oil) based on a calculated 3 ml/kg
dosage for inducing liver injuries according
to lee et al 2005. The yoghurt diet was 180
cc3 per kg body weight per day. The feeding
period continued for 6 weeks. During the
experimental period, the food consumption
and body weight of the rats were followed.
The values of the biochemical analysis were
expressed as mean ± SD and ± SEM and
statistically analyzed using one way analysis
of variance (ANOVA). Student t test was
used for significance. Differences were
considered significant at p<0.05 according
to Artimage and Berry (1987).
187
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
et al 2011; Com and Hisil 2010; Negi et al
2003; Gil et al 2000).
Moreover Turfan et al (2012) reported that
the antioxidative properties of pomegranate
products are correlated with their
polyphenolic content including tannins and
anthocyannins. The rate of increase in TPC
in fresh yoghurt supplemented with 4% and
8% (PJ), Table (2) was 27.7% and 103.6%
respectively, while it was 19.9% and 92.8%
in the same order after 3 days of storage.
During cold storage the TPC gradually
decreased for all samples, this may be
attributed to the transformation of phenolic
compounds which unstable and undergo
numerous enzymatic and chemical reactions
during storage as stated by (Cheynier
(2005); Poncet-Legrand et al (2005); ESSafi et al (2007). Uzuner at al (2011) and
Turfan et al (2012) found that anthocyanins
in pomegranate lost its stability during
storage and some of polyphenols formed diesters with -D-glucose.
Results and Discussion
Textural properties
Table (1) presents the results of texture
analysis
performed
on
yoghurt
supplemented with pomegranate juice (PJ)
and the control sample. Data revealed that
hardness of yoghurt samples was not
significantly affected with increasing the
concentration of (PJ) and storage periods
until 6% then decreased significantly with
increasing the (PJ) added (8%, 10%) and at
the end of storage (12 days). This may be
attributed to the moisture content of (PJ) at
higher concentration which weakens the
protein network resulting in a less firmness
(Ahmed et al 2005). In yoghurt, the protein
matrix consists of short interconnected
chains; consequently the liquid phase of
juice is immobilized in the interstitial spaces
in the protein matrix (Shiby and Mishra
2012). Values of cohesiveness were
relatively lower for yoghurt samples
supplemented with (PJ) than in plain
yoghurt samples, this may due to the
strength of protein-protein interaction bonds
in control yoghurt rather that in the mixture
of milk and (PJ) which weakened this
phenomena. Also, noticed the same results
in springiness data. Moreover, chewiness
and gumminess of the yoghurt supplemented
with (PJ) were reduced significantly rather
than control yoghurt (without PJ).
Antioxidant activity (AA)
DPPH radical scavenging activity (RSA) of
yoghurt supplemented with (PJ) had higher
antioxidant activity than control yoghurt
(without juice) and this was directly
proportional to the percentage of added juice
Fig (2). This ascribed to the phenolic
compounds found in pomegranate which are
responsible for 92% of its AA (Seeram et al
2006 and Kulkarni et al 2004). The rate of
increase in RSA by adding 4% and 8%
pomegranate was 198% and 398 %
respectively for fresh yoghurt samples,
Table (3), while the corresponding rates
after 3 days of storage was 106.5% and
321.7%. After 12 days of storage, the rate of
increase in RSA for 4% and 8%
concentration of pomegranate were 28.6%
and 125.7% respectively.
Total phenolic content (TPC):
Results illustrated in Fig. (1) revealed that
TPC of yoghurt fortified with (PJ) increased
with increasing the concentration from 2%
to 10% this ascribed to the TPC of (PJ)
(3425 gallic acid mg/l). Many researchers
reported that (PJ) is a very rich source of
polyphenols including ellagitannins (Uzuner
188
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
Table.1 Texture analysis performed for yoghurt supplemental
with pomegranate and the control sample
Analysis
Days
0%
2%
4%
6%
8%
10%
hardness
fresh
3 days
1.0
1.0
1.0
1.0
1.0
1.0
1.1
1.0
0.7
0.7
0.6
0.6
6 days
1.0
1.0
1.0
1.0
0.6
0.5
12 days
0.8
0.8
1.0
1.0
0.6
0.5
Cohesivness
Spinginess
Gumminess
Chewiness
fresh
0.82
0.416
0.488
0.404
0.339
0.513
3 days
0.846
0.424
0.496
0.399
0.343
0.516
6 days
0.813
0.419
0.437
0.438
0.429
0.506
12 days
0.641
0.435
0.492
0.589
0.468
0.431
fresh
0.649
0.469
0.495
0.515
0.499
0.583
3 days
0.647
0.469
0.492
0.582
0.514
0.524
6 days
0.67
0.448
0.655
0.561
0.504
0.534
12 days
0.682
0.601
0.642
0.669
0.552
0.532
fresh
0.82
0.416
0.586
0.515
0.237
0.308
3 days
0.846
0.424
0.595
0.399
0.24
0.309
6 days
0.813
0.419
0.568
0.482
0.258
0.253
12 days
0.852
0.479
0.541
0.589
0.281
0.216
fresh
0.532
0.195
0.29
0.265
0.118
0.179
3 days
0.544
0.199
0.293
0.232
0.123
0.162
6 days
12 days
0.544
0.581
0.188
0.288
0.372
0.347
0.27
0.512
0.13
0.155
0.135
0.115
Fig.1 Total phenolic content equivalent gallic acid (mg/l) of yoghurt samples supplemented
with pomegranate and storage in refrigerator at 6±2°C
25
20
15
10
8
5
4
0
12
9
6
3
0
Fresh
Total phenolic content
equivalent gallic acid (mg/l
30
Days
189
Conc.
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
Table.2 Percentage rate of increase in total phenolic content equivalent gallic acid (mg/l) of yoghurt
samples supplemented with pomegranate and storage in refrigerator at 6±2°C
Conc. of
pomegranate
2
4
6
8
Fresh
14.458
24.711
95.783
103.614
3
10.843
14.880
87.952
92.771
Storage periods (days)
6
6.580
19.079
20.395
46.053
9
7.576
27.273
36.364
50.758
12
2.941
8.823
9.804
46.078
Table.3 Rate of increase in DPPH-radical scavenging activity (RSA) for yoghurt supplemented with
pomegranate and storage in refrigerator at 6±2°C
Conc. of
pomegranate
2
4
6
8
Fresh
38.776
194.959
255.102
394.959
3
45.652
106.522
243.478
321.739
Storage periods (days)
6
9
48.780
25.641
58.536
38.462
60.976
43.590
260.976
192.308
12
17.143
28.571
48.571
125.714
Table.4 The weight gain, food intake, liver weight and antioxidant enzyme
(Glutathione & Glutathione peroxidase)
variables
Groups
CRange
Mean ±SD
Mean ± SE
C+
Range
Mean ±SD
Mean ±SE
Y0
Range
Mean ±SD
Mean ± SE
Y4
Range
Mean ± SD
Mean ± SE
Y8
Range
Mean ±SD
Mean ± SE
Weight gain
(g)
Food intake
(g/day)
Liver weight
(g)
(25.00 - 49.00)
32.80±9.39
32.80 ± 4.20a
(12.00- 14.00)
12.80 ±0.84
12.80 ±0.37a
(5.00 - 7.60)
6.18 ±0.93
6.18 ±0.41a
(25.00 31.00)
28.33 ±3.05
28.33 ±1.76a
(250.00 -280.00)
265.00 ± 15.00
265.00 ± 8.66a
(25.00 28.00)
26.20 ±1.30
26.20 ±0.58b
(7.00 9.00)
8.20 ±0.84
8.20 ±0.37c
(5.40 - 6.70)
5.80 ±0.52
5.80 ±0.23a
(19.00 -27.00)
22.00 ±4.36
22.00 ±2.52a
(215.00 240.00)
228.33 ±12.58
228.33 ±7.26b
(26.00 32.00)
28.60 ±2.40
28.6 ±1.07ab
(8.00 10.00)
9.00 ± 0.71
9.00 ± 0.32bc
(5.80 - 6.60)
6.14 ±0.31
6.14 ±0.14a
(23.00 34.00)
28.00 ±5.59
28.00 ±2.21a
(210.00 -260.00)
236.66 ± 25.16
236.33 ±14.53ab
(27.00 32.00)
29.80 ±1.92
29.80 ±0.86ab
(9.00 - 11.00)
10.00 ±1.00
10.00 ±0.45b
(6.00 - 6.80)
6.32 ±0.31
6.32 ±0.14a
(18.00 26.00)
22.33 ±4.04
22.33 ±2.33a
(220.00 -270.00)
245.00 ±25.00
245.00 ±14.43ab
(28.00 36.00)
31.80 ±3.03
31.80 ±1.36ab
(9.00 11.00)
10.00 ±1.00
10.00 ±0.13a
(6.10 - 6.80)
6.50 ±0.29
6.50 ±0.13a
(22.00- 29.00)
25.66 ± 3.51
25.66 ±2.02a
(235.00 -260.00)
250.00 ±13.23
250.00 ±7.64ab
abc
GSH
(mmol/g )
Means in each column having the same letter were not significantly different.
Values with different superscript letters are significantly different at p<0.05.
190
GPx
( /mg)
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
Fig.2 DPPH- radical scavenging activity (RSA) for yoghurt supplemented with pomegranate
and storage in refrigerator at 6±2°C
30
% RSA
25
20
15
10
8
5
4
0
Conc.
12
9
6
3
Fresh
0
Days
Table.5 Liver biomarkers (GOT, GPT, BILE total, BILE direct and Albumin)
Variables
AST
( /ml)
ALT
( /ml)
T.Bilirubin.
( /ml)
D.Bilirubin
( /ml)
Groups
CRange
(16.00 21.00) (14.00 21.00) (0.25 0.54)
(0.01 0.10)
Mean ± SD 18.60 ± 2.52
17.33 ± 3.51
0.37 ± 0.11
0.05 ± 0.05
Mean ± SE
18.60 ± 1.45b
17.33 ± 2.03a
0.37 ± 0.06a
0.05 ± 0.03a
C+
Range
(21.00 29.00) (19.00 23.00) (0.42 0.53)
(0.01 0.18)
Mean ± SD 25.60 ± 4.16
21.00 ± 2.00
0.48 ± 0.05
0.09 ± 0.09
Mean ± SE
25.60 ± 2.40a
21.000 ± 1.15a
0.48 ± 0.03a
0.09 ± 0.05a
Y0
Range
(23.00 27.00) (17.00 23.00) (0.30 0.60)
(0.01 0.15)
Mean ± SD 25.00 ± 2.00
20.33 ± 3.05
0.44 ± 0.15
0.06 ± 0.08
Mean ± SE
25.00 ± 1.15a
20.33 ± 1.76a
0.44 ± 0.08a
0.06 ± 0.05a
Y4
Range
(19.00 -26.00)
(16.00 21.00) (0.24 0.70)
(0.01 0.09)
Mean ± SD 23.00 ± 3.61
18.33 ± 2.52
0.43 ± 0.24
0.06 ± 0.04
ab
a
a
Mean ± SE
23.00 ± 2.08
18.33 ± 1.45
0.43 ± 0.14
0.06 ± 0.03a
Y8
Range
(18.00 23.00) (15.00 19.00) (0.26 0.59)
(0.01 - 0.15)
Mean ± SD 20.60 ± 2.52
16.66 ± 2.08
0.39 ± 0.18
0.06 ± 0.08
ab
a
a
Mean ± SE
20.60 ± 1.45
16.66 ± 1.20
0.39 ± 0.10
0.06 ± 0.05a
abc
Means in each column having the same letter were not significantly different.
Values with different superscript letters are significantly different at p<0.05.
191
Albumin
(g/dl)
(3.50 - 3.80)
3.63 ± 0.15
3.63 ± 0.09ab
(3.40 3.50)
3.43 ± 0.05
3.43 ± 0.03b
(3.60 3.90)
3.76 ± 0.15
3.76 ± 0.09a
(3.50 - 3.80)
3.63 ± 0.15
3.63 ± 0.09ab
(3.60 3.80)
3.70 ± 0.10
3.70 ± 0.06a
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
However, the percentage of RSA
gradually decreased during cold storage
for all samples, this presumably due to the
decrease stability of phenolic compounds
and anthocyannin pigments during storage,
this finding was agreement with Sagdic et
al (2012); Mirsaeedghazi et al (2011).
higher mean value (32.80± 4.2g) than
other four groups, followed by Y8 group
(31.80± 1.36g). Food intake values as
comparable with weight gain values,
showed significant elevation in groups Y4
and Y8 (10±0.45 and 10.00 ± 0.13 g/day
respectively) rather than Y0 (9.00± 0.32
g/day). Also, it was noticed that liver
weight was increased for groups Y4 and
Y8 with mean ± SR 6.32± 0.14g and 6.5±
0.13g respectively, while it was 5.8± 0.23
for group C+.
Biochemical assessments
As expected, the subcutaneous injection of
CC14 into rats is known to cause liver
necrosis, alter various biomarkers such as
increasing serum amino transferase
activity and lipid peroxidation and depress
the antioxidant system in the liver (Lee et
al 2007). A similar action was noticed in
group C+. Table (4) showed that weight
gain value in negative control (C- ) had a
The antioxidant enzyme glutathione in C+
group reduced to 22.33±2.33 mmol/g than
C- group (28.33± 1.76 mmol/g). Moreover
GPX value was significantly increased in
groups feed on yoghurt supplemented with
pomegranate in comparison to C+.
192
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
The effect of pomegranate on GSH level is
due to its polyphenols, where, it is know to
be able to modulate the transcription and
expression of proteins related to the
endogenous antioxidant defense by
interacting with antioxidant response
elements in genre promoter regions of
genes encoding proteins related to
oxidative injury management (Moskaug et
al (2005); Myhrstad et al (2002).
histopathological changes (Fig. 3, Pic. E).
Changes in liver biomarkers values in
Table (5) demonstrated that injection with
CC14, group C+ increased blood AST,
ALT, T. and D. Bilirubin and deceased
Albumin, whereas groups Y4 and Y8
exhibited an ability to protect the
hepatotoxicity by deceasing the level of
AST, ALT and blood bilirubin as well as
deceasing the blood albumin and this was
parallel with the increase of pomegranate
added to yoghurt in Y4 and Y8 group.
Similar results were reported by
Luangpirom et al (2013).
Consequently, supplementing yoghurt
with 8% pomegranate juice had a slight
effect on its textural properties, but had a
high positive effects on its total phenolic
content and antioxidant properties.
Moreover,
the
biochemical
and
histopathological studies evidenced a
remarkable improvement of the hepatic
cells. This confirm the ability of
pomegranate
to
protect
afainis
hepatotoxicity (liver damage) induced by
chemicals.
This ascribed to presence of different
bioactive compounds in pomegranate
mainly polyphenols, falvonoids and
anthocyannins which had antioxidant
properties. Similar results were reported
by Basu and Penugonda (2009); Amal et
al (2012); Pirinccioglu et al (2012); Fatma
et al (2013); Luangpiram et al (2013).
References
Histopathological studies
Adams, L.S.; Seeram, N.P.; Aggarwal,
B.B.; Takada, Y.; Sand, D.; Heber, D.
(2006). Pomegranate Juice, Total
Pomegranate
Ellagitannins,
and
Punicalagin Suppress Inflammatory
Cellsignaling in Colon Cancer Cells.
Journal of Agricultural and Food
Chemistry, 54(3): 980-985.
Adams, L.S.; Zhang, Y.; Seeram, N.P.;
Heber, D.; Chen, S. (2010).
Pomegranate
Ellagitannin-derived
Compounds Exhibit Antiproiferation
and Antiaromatase Activity in Breast
Cancer Cells In vitro. Cancer
Prevention Research, 3(1):108-113.
Ahmed, N.H.; El Soda, M.; Hassan, A.N.;
Frank, J. (2005). Improving the
textural properties of an acidcoagulated (Karish) cheese using
exopolysaccharide producing cultures.
The microscopical examination on liver
section of control rat (C- group) showed a
normal histological structure of hepatic
lobule (Fig. 3, Pic. A), while, liver section
of CC14-treated rats (C+ group) revealed
to the presence of focal hepatic necrosis
associated with inflammatory cells
infiltration (Fig. 3, Pic B). Administration
with yoghurt only (Y0 group) for CC14treated rates, the liver cells still had
vacuolation of hepatocytes, congestion of
hepatic sinusoids and degeneration of
hepatocytes
(Fig.
3,
Pic.
C).
supplementing
yoghurt
with
4%
pomegranate (Y4 group), live section (Fig.
3, Pic. D) Showed cytoplasmic
vacuolization of hepatocytes, while rats
feed on yoghurt supplemented with 8%
pomegranate (Y8 group) showed no
193
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
LWT - Food Science and Technology,
38:843-847.
Amal, A. F.; Abd El-Kader M.A.; Abd ElHaleem A.H. (2012). Modulatory
Effects of Pomegranate Juice on
Nucleic Acids Alterations and
Oxidative Stress in Experimentaly
Hepatitis Rats. Life Science Journal,
9(3):676-682.
Artimage, G.Y.; Berry, W.G. (1987).
Statistical Methods 7th Ed. Ames,
Iowa Stata University Press, 39- 63.
Aswal, P.; Shukla, A.; Priyadarshi, S.
(2012).
Yoghurt
preparation
characteristics
and
recent
advancements Cibtech Journal of BioProtocols 1:32-44.
Bartholomev, R.J.; Delany, A. (1966).
Proc Aust. Assoc .Biochemists., 1:
214.
Basu, A.; Penugonda, K. (2009).
Pomegranate juice: a heart-healthy
fruit juice. Nutr Rev. 67(1):49-56
Cheynier, V. (2005). Polyphenolis in food
are more complex than often though.
Amer. J. of Clin. Nutr., 81:223S229S.
Coïsson, J.D.; Travaglia, F.; Piana, G.;
Capasso, M.; Arlorio, M. (2005).
Euterpe oleracea juice as a functional
pigment for yogurt. Food Research
International, 38:893-897
Com, M.; Hisil, Y. (2010). Pressurized
water extraction of polyphenols from
pomegranate peels. Food Chemistry,
123: 878 885.
Draper, H.H.; Hadley, M. (1990).
Malondialdehyde determination as
index of lipid peroxideation, Methods
Enzymol., 186: 421-431.
Duncan, D B. (1955). Multiple range and
multiple F tests. Biometrics 11:1 42.
Ellman, G.L. (1959). Tissue sulfhydryl
groups. Arch. Biochem. Biophys., 82:
70-77.
Es-Safi, N.; Ghidouche, S.; Ducrot, P.H.
(2004). Flavonoids: Hemisynthesis,
Reactivity, Characterization and Free
Radical
Scavenging
Activity.
Molecules 2007, 12, 2228-2258.
Fatma, E.A.; Hassannane, M.M; Omara,
E.A; Hasan A.M.; El-Toumy, S.A.
(2013). Protective Effect of Punica
granatum Peel Extract Against
Pentachlorophenol-Induced Oxidative
Stress, Cytogenetic Toxicity and
Hepatic Damage in Rats. Australian
Journal of Basic and Applied
Sciences, 7(2): 853-864,
Ghadge, P.N.; Prasad, K.; Kadam, P.S.
(2008). Effect of fortification on the
physico-chemical
and
sensory
properties of buffalo milk yoghurt.
Electr. J. Environ. Agric. Food Chem.,
7: 2890-2899.
Gil, M.I.; Tomas-Barberán, F.; HessPierce, B.; Holcroft, D.; Kader, A.
(2000). Antioxidant activity of
pomegranate juice and its relationship
with phenolic composition a d
processing. J. Agric. Food Chem.
48:4581-4589.
Heber,
D.
(2011).
Pomegranate
Ellagitannins. In: Herbal medicine
Biomolecular and Clinical Aspects.
2nd edition. Benzie.I.F.F., wachtelGalor S. (eds). Chapter 10, Boca
Raton (F1):CRC Press, USA.
International Dairy Federation. (1991).
Rheological and Fracture Properties
of Cheese. Brussels, Belgium:
International Dairy Federation;Bull.
IDF No. 268
Kalkarni, A.P.; Aradhya, S.M.; Divakar, S.
(2004). Isolation and identification of
a radical scavenging antioxidant
punicalagin from pith and carpellary
membrane of pomegranate fruit. Food
chem. 84:551-557.
Kaneria, M.J.; Bapodara, M.B.; Chanda,
S.V. (2012). Effect of extraction
techniques
and
solvents
on
194
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
antioxidant activity of pomegranate
(Punica granatum L.) leaf and stem.
Food Anal. Methods, 5: 396-404.
Kulkarni, A.P.; Aradhya, S.M.; Divakar,
S. (2004). Isolation and identification
of a radical scavenging antioxidant
punicalagin from pith and carpellary
membrane of pomegranate fruit. Food
Chemistry, 87:551-557.
Lee, C.P.; Shih, P.H.; Hsu, C.L.; Yen,
G.C. (2007). Hepatoprotection of tea
seed oil (Camellia oleifera Abel.)
against CCl4 induced oxidative
damage in rats. Food Chem Toxicol.,
45: 888 95.
Lee, G.P.; Jeong, W.I.; Jeong, D.H.; Do,
S.H.; Kim, T.H.; Jeong, K.S. (2005).
Diagnostic evaluation of carbon
tetrachloride-induced rat hepatic
cirrhosis model. Anticancer Res.,
Mar-Apr; 25(2A): 1029-38.
Lourens-Hattingh, A.; Viljoen, B.C.
(2001). Review: Yoghurt as probiotic
carrier in food. Int. Dairy J. 11: 1-17
Luangpirom, A.; Junaimuang, T.;
Kourchampa, W.; Somsapt, P.;
Sritragool, O. (2013). Protective effect
of pomegranate (Punica granatum
Linn.) juice against hepatotoxicity and
testicular toxicity induced by ethanol
in mice. ABAH Bioflux, 5(1):87-93.
Meydani, S.N.; Ha, W. (2000).
Immunologic effects of yogurt. Am J
Clin Nutr.71:861 72.
Mirsaeedghazi, H.; Emam-Djomeh, Z.;
Ahmed K.R. (2011).Effect of frozen
storage on the anthocyannins and
phenolic compounds of pomegranate
juice. Journal of food science and
technology 23:11-35.
Moskaug, J.O.; Carlsen, H.; Myhrstad,
M.C.;
Blomhoff,
R.
(2005).
Polyphenols and glutathione synthesis
regulation. Am. J. Clin. Nutr., 81:
277S-283S.
Myhrstad, M.C.; Carlsen, H.; Nordstrom,
O.; Blomhoff, R.; Moskaug, J.O.
(2002). Flavonoids increase the
intracellular glutathione level by
transactivation
of
the
gamma
glutamylcysteine
synthetase
catalytical subunit promoter. Free
Radic. Biol. Med., 32: 386-393.
Negi, P. S.; Jayaprakasha, G. K.; Jena, B.
S.
(2003).
Antioxidant
and
antimutagenic
activities
of
pomegranate peel extracts. Food
Chemistry, 80, 393 397.
Pirinccioglu, M.; Kizil, G.; Kizil, M.;
Kanay, Z.; Ketani, A. (2012). The
protective role of pomegranate juice
against carbon tetrachloride-induced
oxidative stress in rats. Toxicol Ind
Health 16:1-9.
Poncet-Legrand, C.; Edelmann, A.;
Putaux, J.L.; Cartalade, D.; SarniManchado, P.; Vernhet A. (2005).
Poly (L-proline) interactions with
flavon-3-ols Units: Influence of the
molecular
structure
and
the
polyphenol/protein
ration.
Food
Hydrocolloids, 20(5):677-387.
Reitman, S.; Frankel, S. (1957).
Determination of glutamate pyruvat
transaminase
and
glutamate
oxaloacetate transaminase. Amer. J.
Clin. Path., 28: 56-63.
Ross, M.H.; Reith, E.J.; Rombell, L.J.
(1989). Histology: A text and Atlas
(2nd ed). Battimore: Williams &
Wilkins
Sagdic, O.; Ozturk, I.; Cankurt, H.;
Tornuk, F. (2012). Interaction
between some phenolic compounds
and probiotic bacterium in functional
ice cream production. Food and
Bioprocess Technology, 5(8):29642971
SAS (1998). Statistical analysis system
SAS user's Guide; statistic SAS
institute INC., cory NC.
Seeram, N.P.; Zhang, Y.; Reed, J;
195
Int.J.Curr.Microbiol.App.Sci (2014) 3(8) 185-196
Krueger, C.; Vaya, J.
(2006).
Pomegranate phytochemicals. Chapter
1, pp. 3-30. In: Pomegranates:
Ancient Roots Modern Medicine.
Seeram, N.P.; Schulman, R.N.; Heber,
D. (Eds). CRC Press & Taylor and
Francis Group, Medicinal & Aromatic
Plant Series, Boca Raton, FL, USA.
101, 1365 1371.
Zheng, H. H.; Tu, P. F.; Zhou, K.; Wang,
H.; Wang, B. H. and Lu, J. F. (2001).
). Antioxidant properties of phenolic
diterpenes
from
Rosmarinus
officinalis Acta Pharmacol Sin.
;22(12):1094
Shiby, V.K.; Mishra, H.N. (2012). Effect
of starter culture level on textural and
sensory properties of buffalo milk
Dahi (curd). Egyptian Journal of
Dairy Science. 40(1):15-23
Tomás-Barberán, F.A.; Seeram, N.P.;
Espín, J.C. (2006). Bioavailability of
pomegranate polyphenols. In: Seeram
NP, Schulman RN, Heber D, editors.
Pomegranates. Ancient Roots to
Modern Medicine. Chapter 3. Boca
Raton, FL, USA: CRC Press; 45 60.
Turfan, O. ; Turkyilmaz M. ; Yemis, O.
and Ozkam M.(2012). Effects of
clarification
and
storage
on
anthocyanin and color of pomegranate
juice concentrates. J of Food Quality
vol. 35 issue 4 page: 242-282.
Uzuner, S.; Onsekizoglu, P.; Acar, J.
(2011).
Effects
of
processing
techniques and cold storage on ellagic
acid concentration and some quality
parameters
of
pomegranate
juice.GIDA , Journal of Food. 36 (5):
263-269
Weiss, C.; Marker H.S.; Lehrer, G.M.
(1980). Sensitive fluorometric assays
for glutathione peroxidase and
reductase. Anal Biochem, 106: 512516.
Xuewu, D.; Yueming, J.; Xinguo, S.;
Zhaoqi, Z. and John, S. (2007).
Antioxidant
properties
of
anthocyanins extracted from litchi
(Litchi chinenesis Sonn.) fruit
pericarp tissues in relation to their role
in the pericarp browning. Food Chem.
196